鸭源鸡杆菌ompW基因缺失菌株的构建及其药物敏感性分析

发布时间：2018-06-19 16:46

本文选题：鸭源鸡杆菌 + OmpW　；参考：《畜牧兽医学报》2017年07期

【摘要】：探索OmpW在鸭源鸡杆菌耐药性形成中的作用。运用M-IV培养基诱导鸭源鸡杆菌自然感受态细胞,用含有氯霉素抗性(Cmpr)筛选标记和同源臂的线性DNA打靶片段替换目的基因;以PCR、SDS-PAGE及Western blot鉴定阳性转化子ΔompW。通过微量稀释法测定12种抗菌药物对鸭源鸡杆菌亲本菌株RU和突变株ΔompW的最小抑菌浓度(MIC)。结果显示:鸭源鸡杆菌阳性转化子缺失了ompW,成功构建了ompW缺失突变菌株ΔompW;相对于RU,土霉素和四环素对ΔompW的MIC值分别降低到1/8(16/128,P0.01)和1/32(4/128,P0.01),其他药物的MIC值无明显变化。本研究首次利用自然转化法构建了鸭源鸡杆菌ompW缺失菌株ΔompW;OmpW在鸭源鸡杆菌抗四环素类药物中发挥重要作用。
[Abstract]:To explore the role of OmpW in the formation of drug resistance of chicken bacillus from duck. M-IV medium was used to induce the natural receptive cells of Chicken from Duck, the target gene was replaced by the linear DNA targeting fragment containing chloramphenicol resistant Cmpr1, and the positive transformant 螖 ompW was identified by PCR SDS-PAGE and Western blot. The minimal inhibitory concentration (MIC) of 12 antimicrobial agents against RU and 螖 ompW was determined by microdilution method. The results showed that ompW was absent from the positive transformants of Bacillus duckii, and the ompW deletion mutant 螖 ompW was successfully constructed, and the MIC values of oxytetracycline and tetracycline to 螖 ompW decreased to 1 / 816 / 128 / 128P0.01 and 1 32 / 4 / 128P0.01, respectively, and the MIC values of other drugs did not change significantly. In this study, the ompW deletion strain 螖 ompWOmpW was constructed by natural transformation for the first time to play an important role in the tetracycline resistance of Duck origin chicken bacilli.
【作者单位】：河南农业大学禽病研究所;中国兽医药品监察所;
【基金】：国家自然科学基金(31302090) 河南省自然科学基金(162300410153) 河南省高校科技创新团队与支持计划资助项目(14IRTSHN015)
【分类号】：S852.61

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